Megan L. Uhelski

The anterior cingulate cortex and pain processing

The neural network that contributes to the suffering which accompanies persistent pain states involves a number of brain regions. Of primary interest is the contribution of the cingulate cortex in processing the affective component of pain. The purpose of this review is to summarize recent data obtained using novel behavioral paradigms in animals based on measuring escape and/or avoidance of a noxious stimulus. These paradigms have successfully been used to study the nature of the neuroanatomical and neurochemical contributions of the anterior cingulate cortex (ACC) to higher order pain processing in rodents.

Maternal separation stress leads to enhanced emotional responses to noxious stimuli in adult rats

The purpose of the current study was to examine pain processing in adult rats following repeated maternal separation in infancy, a common model of early life stress. Sensory pain processing remained unaltered, as measured using threshold testing of nociception. However, affective pain processing was enhanced as revealed by increased responding during the tonic phase of the formalin test and during the place escape/avoidance test. The pattern of enhanced responses suggests that early life stress alters the emotional response to pain. Further research could determine if this pattern holds true for different pain models, or if post-weaning enrichment could reverse the effects of maternal separation on pain processing.

Pain inhibition by optogenetic activation of specific anterior cingulate cortical neurons.

Cumulative evidence from both humans and animals suggests that the anterior cingulate cortex (ACC) is important for pain-related perception, and thus a likely target for pain relief therapy. However, use of existing electrode based ACC stimulation has not significantly reduced pain, at least in part due to the lack of specificity and likely co-activation of both excitatory and inhibitory neurons. Herein, we report a dramatic reduction of pain behavior in transgenic mice by optogenetic stimulation of the inhibitory neural circuitry of the ACC expressing channelrhodopsin-2. Electrophysiological measurements confirmed that stimulation of ACC inhibitory neurons is associated with decreased neural activity in the ACC. Further, a distinct optogenetic stimulation intensity and frequency-dependent inhibition of spiking activity in the ACC was observed. Moreover, we confirmed specific electrophysiological responses from different neuronal units in the thalamus, in response to particular types of painful stimuli (i,e., formalin injection, pinch), which we found to be modulated by optogenetic control of the ACC inhibitory neurons. These results underscore the inhibition of the ACC as a clinical alternative in inhibiting chronic pain, and leads to a better understanding of the pain processing circuitry of the cingulate cortex.

Pain affect in the absence of pain sensation: evidence of asomaesthesia after somatosensory cortex lesions in the rat.

Summary Electrolytic lesions of the hind limb area of the primary somatosensory cortex attenuated paw withdrawal threshold responses without altering pain affect in adult female rats. Abstract Multidimensional models of pain processing distinguish the sensory, motivational, and affective components of the pain experience. Efforts to understand underlying mechanisms have focused on isolating the roles of specific brain structures, including both limbic and non‐limbic cortical areas, in the processing of nociceptive stimuli. The purpose of this study was to examine the role of the somatosensory cortex in both sensory and affective aspects of pain processing. It was hypothesized that animals with lesions of the hind limb area of the somatosensory cortex would demonstrate altered sensory processing (asomaesthesia, a deficit in the ability to detect and identify somatic sensation) in the presence of an inflammatory state when compared to animals with sham lesions. The level of pain affect produced by an inflammatory pain condition was not expected to change, as this region has not demonstrated a role in processing the affective component of pain. Seventy‐nine adult female Sprague‐Dawley rats were randomly assigned to receive bilateral lesions or a sham procedure. The results showed that somatosensory lesions to the hindlimb region altered responses to mechanical stimulation in the presence of experimentally‐induced inflammation, but did not attenuate the inflammation‐induced paw volume changes or the level of pain affect, as demonstrated by escape/avoidance behavior in response to mechanical stimulation. Overall, these results support previous evidence suggesting that the somatosensory cortex is primarily involved in the processing the sensory/discriminative aspect of pain, and the current study is the first to demonstrate the presence of pain affect in the absence of somatosensory processing.

Impairment of Recovery From Incentive Downshift After Lesions of the Anterior Cingulate Cortex: Emotional or Cognitive Deficits?

The anterior cinculate cortex (ACC) is known to be implicated in pain-fear and reward expectations. Animals were given electrolytic lesions of the ACC and then trained in the consummatory successive negative contrast (cSNC) situation. In cSNC, animals exposed to an incentive downshift from 32% to 4% sucrose exhibit less consummatory behavior than animals always exposed to 4% sucrose. The ACC lesion had no measurable effects on the consummatory performance of animals before the downshift (i.e., the lesion did not affect consumption of 32% vs. 4% sucrose); on the performance of unshifted, 4% sucrose animals; and on the first downshift trial. However, ACC animals exhibited a significant retardation of recovery from cSNC relative to downshifted shams. Within-trial analysis of consummatory behavior indicated that ACC lesions facilitated cSNC during both the initial and last 100 s of postshift trials after the first downshift experience, relative to sham controls. These results suggest that the ACC is part of the neural circuit normally involved in coping with the emotional response induced by the incentive downshift event by inducing learning of the new incentive conditions.

Hyperbaric oxygen treatment decreases pain in two nerve injury models

Hyperbaric oxygen (HBO) treatment has been used clinically to treat a variety of ailments, including severe burns and carbon monoxide poisoning, and in research settings has produced promising results when used to treat animal models of inflammatory pain. However, studies examining neuropathic pain or nerve injury models have been limited to physiological assessments and not whether the pain condition improves. The purpose of this study was to evaluate the effect of HBO on two common models of neuropathic pain, L5 ligation and chronic constriction injury (CCI) of the sciatic nerve. Following surgical manipulations, animals demonstrating mechanical hyperalgesia were randomly assigned to either HBO treatment or control for 90min treatment sessions, after which mechanical sensitivity was assessed at 15min and 6h post. Daily HBO sessions, with assessments 15min post-treatment, continued for two weeks, followed by 5 days of assessment only. The results indicated that both models demonstrated significant improvement in response to treatment over the course of the two-week period, with CCI animals recovering more quickly and maintaining this recovery throughout the post-treatment period. Hyperbaric oxygen treatment appears to be successful in relieving neuropathic pain for an extended period of time, and future research should be aimed at investigating the precise mechanisms underlying this positive effect.

Evaluating underlying neuronal activity associated with escape/avoidance behavior in response to noxious stimulation in adult rats

The place escape/avoidance paradigm (PEAP) is a behavioral test designed to quantify the level of unpleasantness evoked by painful stimuli by assessing the willingness of a subject to escape/avoid a preferred area when it is associated with noxious stimulation. Previous studies have demonstrated that escape/avoidance behavior is dependent on activity in the anterior cingulate cortex (ACC), a region of the limbic system involved in processing the emotional component of pain in humans and animals. Analysis of c-Fos expression in the ACC confirmed that the escape/avoidance response to noxious stimuli corresponds to changes in neural activation in this region. Behavioral tests such as the PEAP may be more sensitive to changes in supraspinal pain processing and could contribute to the development of novel analgesics in the future.

Examining the role of the medial thalamus in modulating the affective dimension of pain

The purpose of this project was to explore the role of the medial thalamus (MT), including the medial dorsal thalamus (MD) and associated midline nuclei in pain processing. Experiment 1 explored the role of electrolytic lesions to the MT in the formalin test. It was hypothesized that animals with electrolytic lesions to the MT would have attenuated paw licking behavior during the second phase of the formalin tests as compared to sham lesion controls. This hypothesis was based on evidence of projections from the MD to the ACC, and previous research demonstrating attenuation of paw licking behavior in the second phase of the formalin test in animals with ACC lesions. Experiment 2 tested the effects of electrolytic MT lesions on mechanical paw withdrawal thresholds in the L5 nerve ligation model. It was hypothesized that lesions of the MT would not alter mechanical paw withdrawal thresholds. Experiment 3 tested the effects of electrolytic MT lesions on escape/avoidance behavior in the place escape avoidance paradigm. For experiment 1, animals with MT lesions were found to have slightly elevated paw licking behavior, but only across two time points. No differences in mechanical paw withdrawal thresholds and in escape/avoidance behavior were detected as compared to the sham lesion group. These results indicate a limited role for the medial thalamic nuclei in coding for pain intensity and the affective dimension of pain. Additional research is needed to explore the role of individual medial nuclei in pain processing.

Inhibition of anandamide hydrolysis attenuates nociceptor sensitization in a murine model of chemotherapy-induced peripheral neuropathy

Painful neuropathy frequently develops as a consequence of commonly used chemotherapy agents for cancer treatment and is often a dose-limiting side effect. Currently available analgesic treatments are often ineffective on pain induced by neurotoxicity. Although peripheral administration of cannabinoids, endocannabinoids, and inhibitors of endocannabinoid hydrolysis has been effective in reducing hyperalgesia in models of peripheral neuropathy, including chemotherapy-induced peripheral neuropathy (CIPN), few studies have examined cannabinoid effects on responses of nociceptors in vivo. In this study we determined whether inhibition of fatty acid amide hydrolase (FAAH), which slows the breakdown of the endocannabinoid anandamide (AEA), reduced sensitization of nociceptors produced by chemotherapy. Over the course of a week of daily treatments, mice treated with the platinum-based chemotherapy agent cisplatin developed robust mechanical allodynia that coincided with sensitization of cutaneous C-fiber nociceptors as indicated by the development of spontaneous activity and increased responses to mechanical stimulation. Administration of the FAAH inhibitor URB597 into the receptive field of sensitized C-fiber nociceptors decreased spontaneous activity, increased mechanical response thresholds, and decreased evoked responses to mechanical stimuli. Cotreatment with CB1 (AM281) or CB2 (AM630) receptor antagonists showed that the effect of URB597 was mediated primarily by CB1 receptors. These changes following URB597 were associated with an increase in the endocannabinoid anandamide in the skin. Our results suggest that enhanced signaling in the peripheral endocannabinoid system could be utilized to reduce nociceptor sensitization and pain associated with CIPN.

Role of the ventrolateral orbital cortex and medial prefrontal cortex in incentive downshift situations

The present research evaluated the role of two prefrontal cortex areas, the ventrolateral orbital cortex (VLO) and the medial prefrontal cortex (mPFC), on two situations involving incentive downshifts, consummatory successive negative contrast (cSNC) with sucrose solutions and Pavlovian autoshaping following continuous vs. partial reinforcement with food pellets. Animals received electrolytic lesions and then were tested on cSNC, autoshaping, open-field activity, and sucrose sensitivity. Lesions of the VLO reduced suppression of consummatory behavior after the incentive downshift, but only during the first downshift trial, and also eliminated the enhancement of anticipatory behavior during partial reinforcement, relative to continuous reinforcement, in autoshaping. There was no evidence of specific effects of mPFC lesions on incentive downshifts. Open-field activity was also reduced by VLO lesions, but only in the central area, whereas mPFC lesions had no observable effects on activity. Animals with mPFC lesions exhibited decreased consumption of the lowest sucrose concentration, whereas no effects were observed in animals with VLO lesions. These results suggest that the VLO may exert nonassociative (i.e., motivational, emotional) influences on behavior in situations involving incentive downshifts. No clear role on incentive downshift was revealed by mPFC lesions.